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Gene Synthesis> | … inally synthesized with the native PglnA promoter/RBS upstream of the sf-gfp gene and with the LVA degradation tag and double terminator downstream of the gene in the pUC57 plasmid (GenScript, Inc., NJ). The complete gene was clone at the XbaI-HindIII site in pACYC184 6 … | Get A Quote |
While predictable design of a genetic circuit's output is a major goal of synthetic biology, it remains a significant challenge because DNA binding sites in the cell affect the concentration of available transcription factors (TF). To mitigate this problem, we propose to use a TF that results from the (reversible) phosphorylation of protein substrate as a circuit's output. We demonstrate that by comparatively increasing the amounts of substrate and phosphatase, the TF concentration becomes robust to the presence of DNA binding sites and can be kept at a desired value. The circuit's input/output gain can, in turn, be tuned by changing the relative amounts of the substrate and phosphatase, realizing an amplifying... More